Figure 4.

Silencing of miR-146a promotes osteoclast differentiation in Raw264.7 cells. (A) Study design of the experiment. Cells were treated with 200 nM miR-146a inhibitor for 24 h, then 30 ng/ml RANKL and 100 ng/ml LPS were added and incubated for 3 days. (B) The expression levels of miR-146a. (C) Numbers of osteoclasts were determined by averaging the positive cell number of six fields of view under the microscope. Bars represent the mean ± standard error of the mean. (D) Subsequent to culturing, the cells were fixed and stained for TRAP. TRAP+ cells with >3 nuclei were classified as osteoclasts. Magnification, ×200. *P<0.05. miR, microRNA; LPS, lipopolysaccharide; RANKL, receptor activator of nuclear factor κβ ligand; TRAP, tartrate-resistant acid phosphatase; NC, negative control; MNCs, multinucleated cells.